BacPower™ Customized Service

Service Details

Note:
* Custom requirements may extend the turnaround  time.
** Applies for proteins under 500AA

Service Feature - Soluble Protein Strategies

Solution 1
Various Tag Screening

Solution 2
Standard Denaturation

Solution 3
Professional Inclusion Body Refolding Services

• Purpose: Easily express fusion protein in soluble form
• Feature: 11 kinds of fusion tag modifications from High Expression_pET-28 a (+)./In stock functionalized vectors with tag in N-terminal
• Advantages: Time saving without lengthy dialysis or dilution step.

• Purpose: Maintain the soluble state of the protein
• Feature: Re-dissoluble in detergent buffer and dialysis can be provided, which protein may have no activity
• Application: Antigen (generate antibody to western Blot generally)
• Workflow: Expression evaluation > Scale up expression > IBs washing > IBs solubilization >Purification > Dialysis and QC

• Purpose: Eagle to obtain active protein but failed in supernatant or other cells
• Feature: A lot of efforts on refolding condition screening, IB treatment, purification and analysis methods, which can be more close to nature refolding with activity
• Application: For large scale expression and refolding
• Technical effort:
  • 17 kinds of refolding buffer screening by dilution refolding way
  • Check refolding results by reduced and non reduced SDS-PAGE, SEC-HPLC and LC-MS

Case Studies

• Case 1: Professional refolding_Protein

  Background

  • rhBMP9 protein, 3 inner-chain disulfide bond and 1 inter-chain disulfide bond

  Key Challenge

  • Hard to express in soluble condition when expressed in E.coli
  • Up to 2mg/L in CHO cell

  Optimization

  • Inclusion body treatment and denaturation process
  • Refolding buffer screening by dilution refolding way

  Result

  • Final Yield: 73mg/L
  • SEC-HPLC purity: Up to 99%

  

  

  
• Case 2: Enzyme manufacturing
  • We have extensive experience with high-quality enzyme production, including vector/sequence design, upstream and downstream process optimization, customized QCs as well as activity assays.
  • For different stages in enzyme production, from screening and evolution to supply, our solutions can achieve ideal and customized delivery by expressing at high-density in large volumes, providing high purity.
  • Our scientists are dedicated to solving any existing technical barriers, from cloning strategy to protein purification. For in vitro diagnostics and gene and cell therapy applications, we have successfully delivered various enzymes, including Exonuclease III, TelN Protelomerase, Cas9/Cas13a, MAD7, T4 RNA Ligase, DNA polymerase, BsaI, Benzonase, etc.

  

  

  

  
• Case 3: Cytokine_Interleukins-10

  Background

  • A recognized inflammatory and immunosuppressive factor

  Key Challenge

  • Hard to express in soluble condition when expressed in E.coli
  • limited output

  Optimization

  • Proper incubation to get more protein
  • Inclusion body utilization: wash and refolding

  Result

  • Total Amount: 75.15 mg
  • Expression Scale: 50 L
  • Pure and correct folded protein